Thymidine kinase

Abstract

A polynucleotide comprising a nucleotide sequence encoding a thymidine kinase wherein at least one of the nucleotides corresponding to the splice donor site nucleotides is replaced by another nucleotide and wherein the nucleotides of the splice acceptor sites are not altered.

The invention claimed is: 
     
       1. A method of destroying cells comprising (i) introducing into said cells a polynucleotide selected from the group consisting of the TkMut23 polynucleotide as set forth in SEQ ID NO:3, the TkMut24 polynucleotide as set forth in SEQ ID NO:4, and the TkMut234 polynucleotide as set forth in SEQ ID NO:7, and expressing the thymidine kinase encoded by the polynucleotide in said cells; and (ii) simultaneously, separately or sequentially contacting said cells with a non-toxic agent which is converted by the thymidine kinase to a toxic agent, thereby destroying the cells. 
     
     
       2. The method according to  claim 1 , wherein the non-toxic agent is selected from the group consisting of ganciclovir, acyclovir, triflurothymidine, 1-[2-deoxy, 2-fluoro, β-D-arabino furanosyl]-5-iodouracil, ara-A, ara 1, 1-β-D arabino furanosyl thymine, 5-ethyl-2′deoxyuridine, 5-iodo-5′-amino-2,5′-dideoxyuridine, idoxuridine, AZT, AIV, dideoxycytidine, Ara C, and bromovinyl deoxyuridine (BVDU). 
     
     
       3. A method of treating a patient with cells in need of destruction comprising (i) introducing into the patient a polynucleotide selected from the group consisting of the TkMut23 polynucleotide as set forth in SEQ ID NO:3, the TkMut24 polynucleotide as set forth in SEQ ID NO:4, and the TkMut234 polynucleotide as set forth in SEQ ID NO:7, and expressing the thymidine kinase encoded by the polynucleotide in said cells; and (ii) simultaneously, separately or sequentially introducing into the patient a non-toxic agent which is converted by the thymidine kinase to a toxic agent, thereby destroying the cells and treating said patient. 
     
     
       4. The method according to  claim 3 , wherein the non-toxic agent is selected from the group consisting of ganciclovir, acyclovir, triflurothymidine, 1-[2-deoxy, 2-fluoro, β-D-arabino furanosyl]-5-iodouracil, ara-A, ara 1, 1-β-D arabino furanosyl thymine, 5-ethyl-2′deoxyuridine, 5-iodo-5′-amino-2,5′-dideoxyuridine, idoxuridine, AZT, AIV, dideoxycytidine, Ara C, and bromovinyl deoxyuridine (BVDU). 
     
     
       5. A method of treating a patient with cells in need of destruction comprising (i) removing the cells from the patient or donor cells; (ii) introducing into the cells ex vivo a polynucleotide selected from the group consisting of the TkMut23 polynucleotide as set forth in SEQ ID NO:3, the TkMut24 polynucleotide as set forth in SEQ ID NO:4, and the TkMut234 polynucleotide as set forth in SEQ ID NO:7; (iii) introducing the modified cells into the patient; (iv) allowing the cells to express the thymidine kinase encoded by the polynucleotide; and (v) administering to the patient a non-toxic agent which is converted by the thymidine kinase into a toxic agent, thereby destroying the cells and treating said patient. 
     
     
       6. The method according to  claim 5 , wherein the non-toxic agent is selected from the group consisting of ganciclovir, acyclovir, triflurothymidine, 1-[2-deoxy, 2-fluoro, β-D-arabino furanosyl]-5-iodouracil, ara-A, ara 1, 1-β-D arabino furanosyl thymine, 5-ethyl-2′deoxyuridine, 5-iodo-5′-amino-2,5′-dideoxyuridine, idoxuridine, AZT, AIV, dideoxycytidine, Ara C, and bromovinyl deoxyuridine (BVDU). 
     
     
       7. A method of treating graft-versus-host disease in a patient comprising: (i) administering to the patient T-cells genetically engineered to include a polynucleotide selected from the group consisting of the TkMut23 polynucleotide as set forth in SEQ ID NO:3, the TkMut24 polynucleotide as set forth in SEQ ID NO:4, and the TkMut234 polynucleotide as set forth in SEQ ID NO:7, and expressing the thymidine kinase encoded by the polynucleotide in said T-cells; and (ii) administering to said patient, on occurrence of graft-versus-host disease, a non-toxic agent which is converted by the thymidine kinase to a toxic agent thereby killing said genetically engineered T-cells and treating the graft-versus-host disease in said patient. 
     
     
       8. The method according to  claim 7 , wherein the non-toxic agent is selected from the group consisting of ganciclovir, acyclovir, triflurothymidine, 1-[2-deoxy, 2-fluoro, β-D-arabino furanosyl]-5-iodouracil, ara-A, ara 1, 1-β-D arabino furanosyl thymine, 5-ethyl-2′deoxyuridine, 5-iodo-5′-amino-2,5′-dideoxyuridine, idoxuridine, AZT, AIV, dideoxycytidine, Ara C, and bromovinyl deoxyuridine (BVDU).